nothing is imposibble!

I finished my secondary education in SMK Taman Johor Jaya (1) which is ranked number 2 problematic school during my time, and climbed up to number 1 after I'm grad because my super juniors just burned the security guard pondok.

I was actively involved in St. John Ambulance Malaysia then and enjoying school politics as Duty Secretary of SJAM division SMKTJJ1.

Form 6, I have to changed to another secondary school, SMK Permas Jaya, which is a better managed school, but the students there are just too good, they will walk within the line, everyone uniforms are so neat, the toilet is so clean, nobody smoke, no fire burning, no ppl trying to steal the TV from library and the staff and head master are just too good and fair! make me not used to it!

The SJAM here is like a dead organization. With 300 members in the list. Nobody appeared on Saturday Ko-K activity. No proper activity. No proper AGM. As SJAM members, we decided to help them up.

After several meeting and discussion, I found out many things that I thk is possible, is impossible in their mind, esp from Physics class.

This is opposite of my idea that we should try before we say it is not doable, even we failed, we still can learned something from the failure, of course, we need to plan properly.

After so many years, many ppl still worry this and that, aiya, just do it la when u still young, dont scare fail ma...

However, after so many years, I also kena this virus, injected into my mind liao.

Is time to think again and adjust myself.

IF it is not possible, and if I'm not trying hard enough, I will be regret and SJAM of my form 6 school will not be revived.

Just remind myself not to be too EGO, a little bit can la... haha!

Is time to really think about it.

Colleages

Working with molecular biology, you need imagination, unless you got a microscopic eyes that can zoom into molecules. Here are some tools we using.

A spectrometry, to check on the concentration of bacterial cells, protein, as well as DNA by the reflextion of light.

An Autoclaving machine, heat up to 121C for 15mins, 15bar, in order to sterilize culture media, keep our tools from contamination by microbes.

A weight balance, to measure weight for chemicals, media powder.

Water distiller, to produce clean water free of contaminated chemical like nuclease and protease.

A water bath machine, maintain desired temperature for incubation.

A Polymerase Chain Reaction machine, (PCR), amplification of genes.

A UV light detector, to visualize the results of electrophoresis for analysis.

Kubota centrifugal unit, to separate cells, DNAs, proteins @ desired temperature.

Agarose gel electrophoresis, to run the gel in order to know what u get is what u want.

Not what u want? run again then, u just wasted another 60minutes.

There are a lot other useful stuff for molecular biology such as SDS P.A.G.E for protein analysis, crystallization wells and others.

Mistakes

Sometimes, feeling hopeless and demotivated when things go too slow...

1. E. coli Rosetta-gami2(DE3) pET32-phaCcs

not stable, the expression level keep reducing after sometimes.

Revival of glycerol stock and verification ate most of my times.

Production is still OK. Some part in my mind is asking me to retransform the plasmid again.

However current priority is still crystallization(produce protein fast), and my target for crystallization is end of MAY!

2. Own stupid mistakes(thinking not diverse enough)

Purification in July

The purified PhaC protein seems not pure enough, so, I'm thinking to purify the elution again using the regenerated resin and I did.

However, the results of the SDS PAGE analysis is not good, because the PhaC protein cannot bind efficiently to the resin.

I thought is because of the Regenerated resins. Hence, I try new resin (mistake: I missed something very important here when troubleshooting).

For the 3rd purification, I used the elution from the 2nd purification. The result was the same. Binding of the protein is not efficient. Cannot figure out, think and think and think, and finally....I'm feeling stupid.

There are imidazole with concentration of 250mM in the elution that i used for 2nd and 3rd purification!

My large-scale protein production end here. no more protein. Now speeding for protein production again. I want xstal!

Research, Scientist and Laboratory

Is going to be 6 months soon... The number of time that I'm did my research in lab 414 molecular biology laboratory...

Basically, I like researcher's life, is always full of surprises and disappointments. Of course, very tiring and a very hard efforts are required. So far, There are 4 big stages of my project :

Cloning, Expression, Purification and the crystallization.

Cloning

Construct the desired DNA sequence into a suitable expression vector, and transform into a host, mostly E. coli.

In my case, I'm using pET32 expression vector from Novagen.

Transform into E. coli strain Rosetta-gami 2.

This stage done!

However, I'm trying another expression vector, pCold expression vector, and transform into classic of expression host, E. coli strain BL21 pLySs.

On the way!

Expression

To stimulate the bacterial host in order to over express (translate the gene seq on vector into protein) desired protein.

Optimizations were done, mainly on temperature. 15C, 25C, 30C, 37C. IPTG as inducer with concentration of 0.4mM.

Run SDS PAGE to determine the proteins.

Purification

Using TALON resin, going to try gel filtration soon. maybe can play ion exchange le? yohoo!

Others

6X His Protein Staining, Western Blotting, In vitro enzyme assay

Asian Youth Climate Workshop, Bangkok, 2009

The first time to Bangkok.

The first time into a UN meeting.

The first time marching protest for Climate Change.

It's all happen because of 350.org.

NGO fighting for 350ppm of CO2 level in our atmosphere

The most important number in this planet and the upper safe limit for CO2 level according to Scientists.

The workshop is meaningful and we learned some important skills such as media releasing, event organizing.

The most interesting part is the marching towards UN building.

What the Thai police did is protecting the crowd, unlike my country police, they look for the crowd and stop them.

Anyway, it was a fun event, few thousand people march for

a round 1.30hours, drums, banners, world leader masks, traditional costumes, a lot there!

Just received an email from UM friend, is journals that indicate CO2 level of more than 350ppm in the atmosphere is hurting our coral reef by massive bleaching. Current CO2 level is around 387ppm.

350.org is an international grassroots campaign that aims to mobilize a global climate movement united by a common call to action. By spreading an understanding of the science and a shared vision for a fair policy, we will ensure that the world creates bold and equitable solutions to the climate crisis. 350.org is an independent and not-for-profit project.

What is 350? 350 is the number that leading scientists say is the safe upper limit for carbon dioxide in our atmosphere. Scientists measure carbon dioxide in "parts per million" (ppm), so 350ppm is the number humanity needs to get below as soon as possible to avoid runaway climate change. To get there, we need a different kind of PPM-a "people powered movement" that is made of people like you in every corner of the planet.

Hooray! Congratulation dear all!

To all my dear Award of Merit students,

First, I wish to congrats all of you guys did a very good job in yesterday!

Me and Wei Inn were so impressed by you guys! Especially the towing event!

It has been a long time for the training, and almost every training i almost lost my temper.

I have to apologize if I did any 'lost control' behavior or annoying faces to any one of you.

Frankly, is really not easy to train up a bunch of students like you guys to Award level. The gap is quite big. However, the improvement is amazing, awesome and I'm very proud of you guys!

Thanks for all the good and bad memories we all have together, especially my birthday celebration! haha...

Our journey do not end here, is just a beginning, aim for distinction level, then be a good instructor for the future water sapien generations.

I'm so sorry that this time I'm too busy with 350.org and my master. Also, the stupid spotlight causing the pool to close earlier which mean less training time for us.

Although I don't when will the Distinction exam be held, let's dont stop here, fight on and keep up the spirit!

Why I don't like to swim in USM swimming pool?

I think for most of my same kinds, there are less and less fun to swim in the USM pool.

I still can remember how happy am I during first year, to swim in the pool, after stop my training for about 7 years. That's what I check for after I received offer by USM, wow, there is a pool!

During my time as Junior Exco (S n R), we need to arrange events for outings, need to duty as Lifeguards, check the lifeguard room cleanliness, and of course, help monitor the lifeguard disciplines. But second year I'm still enjoy to do all this stuffs, although my results drop under 3.0.

During that time, we all still happy family.

LAst year, during my 3rd year, is a bad year for me.

Being a vice captain, I need to deal more with the higher authority, the pak cik (guarding the entrance), and also members from lifeguards.

When there are too many things come together, I still able to handle it.

But, when too many politics come together, I feel depressed.

Sports should be just sports! but our dear head of sports unit, is doing a great job to stop the development of student sports.

Let's say about the swimming pool.

This is happen recently.

Case 1, USM lifeguard corps

Unit Sukan hired 3 permanent lifeguards to duty at the pool without inform or discuss with USM lifeguard corps. So one day, 3 lifeguards suddenly pop up! and most importantly, they do not know anything about USM pool rules! They were not informed properly by the unit sukan (I dont think they should be called management nor administration unit).

This is a sign of not respecting students body. As being the most active lifeguard corps in Malaysia, Sports unit USM is trying to stop us from growing, ya, good job, USM lifeguard corps is going down. * And the so call permanent lifeguard is just sitting up at the entrance and look from there, teaching swimming while on duty. This is called PROFESSIONAL! Unit sukan really did a good job!

Case 2, Minden fencers

My friend is the founder of minden fencer, Abe Woo.

I still remember how hard to build it up in the starting point, exam month but every sunday we will gather at unit sukan for training to fence!

That time, our coach is Sir Lim Pei Jing.

he did contributed a lot to build this sports in USM.

Now, he resigned from USM. Why?

Sports unit accept another coach by just looking at the letter and never never consult the team.

This is another similar case, never respect others.

What a loss for Minden Fencer.

Don't think what University can do for you, think what you can do for the university!

bullshit!